TY - JOUR A1 - Müller, Larissa A1 - Jakubowski, Norbert A1 - Hayen, H. A1 - Roos, P.H. T1 - Iodination of proteins, proteomes and antibodies with potassium triodide for LA-ICP-MS based proteomic analyses N2 - A fast and mild method for iodine labelling of proteins is presented which is specifically designed and optimized for laser ablation (LA-)ICP-MS based proteomics. Single proteins (lysozyme, bovine serum albumin, cytochrome c and β-casein), whole proteomes (microsomal proteome of rats) and antibodies (anti-bovine casein, anti-bovine serum albumin) can be efficiently iodinated by means of potassium triiodide with minimal losses of antigen properties and antibody binding to iodinated proteins. A comparison with iodination by use of IODO-Beads is presented and it is shown that triiodide labelling is a fast, cheap and less laborious alternative without compromising the analytical figures of merit. PY - 2011 DO - https://doi.org/10.1039/c1ja10090d SN - 0267-9477 SN - 1364-5544 N1 - Geburtsname von Müller, Larissa: Wäntig, L. - Birth name of Müller, Larissa: Wäntig, L. VL - 26 SP - 1610 EP - 1618 PB - Royal Society of Chemistry CY - London AN - OPUS4-24209 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Polatajko, A. A1 - Feldmann, I. A1 - Hayen, H. A1 - Jakubowski, Norbert T1 - Combined application of a laser ablation-ICP-MS assay for screening and ESI-FTICR-MS for identification of a Cd-binding protein in Spinacia oleracea L. after exposure to Cd N2 - We have studied the binding of the toxic element Cd to plant proteins and have used for this purpose spinach (Spinacia oleracea L.) plants treated with 50 µM Cd(II) as a model system. Laser ablation ICP-MS has been applied for the screening of Cd-binding proteins after separation by native anodal polyacrylamide gel electrophoresis (AN-PAGE) and electroblotting onto membranes. The main Cd-carrying protein band was isolated and investigated by nano-electrospray ionization–Fourier transform ion cyclotron resonance (FTICR) mass spectrometry after tryptic digestion. By this procedure, the main Cd-binding protein was identified as ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO). The latter enzyme has been discussed in the literature to be affected in its activity by oxidative stress induced by Cd. However, in this paper it is demonstrated for the first time that RuBisCO directly binds Cd and thus may be directly altered by this toxic element. A commercially available protein standard was used to verify direct binding of Cd(II) to the protein, even without metabolisation. The resulting metal–protein complex was shown to be stable enough to survive AN-PAGE separation and electroblotting. By the use of size exclusion chromatography coupled with ICP-MS it was demonstrated that the RuBisCO protein standard shows similar metal binding properties to Cd. Furthermore, essential elements such as Mn(II), Fe(II) and Cu(II), which are known to possibly replace the RuBisCO activator Mg(II), were investigated in addition to Zn(II). Again, similar binding properties in comparison to the plant protein were observed. PY - 2011 DO - https://doi.org/10.1039/c1mt00051a SN - 1756-5901 SN - 1756-591X VL - 3 IS - 10 SP - 1001 EP - 1008 PB - RSC Publ. CY - Cambridge AN - OPUS4-24963 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jakubowski, Norbert T1 - Short Course: ICP-MS with Sector Field Devices T2 - 2012 Winter Conference on Plasma Spectrochemistry CY - Tucson, AZ, USA DA - 2012-01-09 PY - 2012 AN - OPUS4-25367 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jakubowski, Norbert T1 - Thermo Fisher Scientific Award in Plasma Spectrochemistry T2 - 2012 Winter Conference on Plasma Spectrochemistry CY - Tucson, AZ, USA DA - 2012-01-09 PY - 2012 AN - OPUS4-25368 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jakubowski, Norbert T1 - Novel challenges for elemental analysis T2 - Bruker European Sales meeting CY - Berlin, Germany DA - 2012-04-25 PY - 2012 AN - OPUS4-25950 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Moreno-Gordaliza, E. A1 - Esteban-Fernández, D. A1 - Giesen, Charlotte A1 - Lehmann, K. A1 - Lázaro, A. A1 - Tejedor, A. A1 - Scheler, C. A1 - Canas, B. A1 - Jakubowski, Norbert A1 - Linscheid, M.W. A1 - Gómez-Gómez, M.M. T1 - LA-ICP-MS and nHPLC-ESI-LTQ-FT-MS/MS for the analysis of cisplatin-protein complexes separated by two dimensional gle elctrophoresis in biological samples N2 - A method for the analysis of Pt–protein complexes in biological samples, previously subjected to cisplatin treatment, has been developed. Proteins were separated by gel electrophoresis, and those bound to Pt were detected with high sensitivity by LA-ICP-(SF)-MS. Pt-containing spots were in-gel digested with trypsin, and the peptides produced identified using nHPLC-ESI-LTQ-FT-MS/MS. The influence of protein separation conditions, staining and gel processing prior to laser ablation on Pt–protein bonds preservation have been evaluated using standard proteins incubated with cisplatin. 2-DE separation under non-reducing conditions followed by either Coomassie blue brilliant or silver staining is appropriate for Pt–protein complexes, achieving a good separating resolution of the proteins in biological samples. Direct LA-ICP-MS analysis of glycerol-treated dried gels for Pt–protein monitoring resulted in better sensitivity, more reliable relative Pt signals and a simpler and less time-consuming approach compared to the analysis of blotted membranes. Ablation of gels allowed tackling protein identification of Pt-spots in the remaining non-ablated material in the gel, making it unnecessary to run several gels in parallel for separate Pt detection and protein identification. By using this approach, Pt coordinated to proteins, such as α-2-macroglobulin, transferrin, albumin or hemoglobin, was detected in the serum from a rat treated in vivo with cisplatin after nrSDS-PAGE separation. Furthermore, the first complete LA-ICP-MS metalloprotein contour map in a 2-DE gel has been produced, in this case for the detection of Pt–protein complexes in renal proximal tubule epithelial cells (RPTECs) incubated with cisplatin. Several proteins were identified in those spots containing Pt, which may have a connection with the drug-induced nephrotoxicity mainly affecting this cell type in the kidney. PY - 2012 DO - https://doi.org/10.1039/c2ja30016h SN - 0267-9477 SN - 1364-5544 VL - 27 IS - 9 SP - 1474 EP - 1483 PB - Royal Society of Chemistry CY - London AN - OPUS4-26562 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jakubowski, Norbert T1 - Development of ICP-MS based methods for label free detection of single cells T2 - European Winter Conference on Plasma Spectrochemistry CY - Kraków, Poland DA - 2013-02-09 PY - 2013 AN - OPUS4-27891 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Richter, Janine A1 - Fettig, Ina A1 - Piechotta, Christian A1 - Philipp, Rosemarie A1 - Jakubowski, Norbert T1 - Tributylzinn in Gesamtwasserproben - Entwicklung eines Referenzverfahrens für die EU-Wasserrahmenrichtlinie PY - 2013 SN - 0016-3538 IS - 9 SP - 2 EP - 4 PB - GIT-Verlag CY - Darmstadt AN - OPUS4-29040 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jakubowski, Norbert T1 - Einführung in die anorganische Massenspektroskopie T2 - 46. Jahrestagung der Deutschen Gesellschaft für Massenspektrometrie CY - Berlin, Germany DA - 2013-03-10 PY - 2013 AN - OPUS4-28032 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Jakubowski, Norbert T1 - Interaction of metals with biomolecules: do we have the proper tools? T2 - "Speciation Seminar" Congress Centre CY - Montpellier, France DA - 2012-05-29 PY - 2012 AN - OPUS4-27353 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -